effect of prenatal administration of valproic acid on the survivability and day of hatching of chick embryo

To assess the effects of prenatal administration of valproic acid on the survivability and day of hatching of chick embryo in comparison with age-matched controls. The experimental study was conducted at the Department of Anatomy, Regional Centre of College of Physicians and Surgeons Pakistan, Islamabad, from February 2010 to February 2011. Fertilised chicken eggs were divided into two groups, labelled as experimental group-A and control group-B. Group-A eggs were injected with valproic acid, incubated and hatched. Group-B eggs underwent sham treatment using normal saline. The fully hatched chicks were then evaluated for the day of hatching and survivability, on hatching or on day 22 of incubation whichever was earlier. Outcome was statistically compared with the controls using SPSS 10. The two groups had 30 eggs each. In Group-A 23[76.66%] chicks hatched out, while there were 7[23.33%] dead chicks. In Group-B, 28[93.33%] chicks hatched out and 2[6.66%] were dead. Chicken embryos exposed to valproic acid in ovo showed increased mortality [p<0.001] and delayed hatching [p<0.001]. Prenatal exposure of chick embryos to valproic acid decreased embryo survival and also delayed hatching compared to age-matched controls

Atrial and ventricular septal changes in ethanol vapour exposed chick embryos

To study the effects of ethanol vapour exposure on development of atrial and ventricular septa of chick embryo. The experimental study was conducted at the College of Physicians and Surgeons, Islamabad, from 2006 to 2007. The experimental and control groups were further divided into three subgroups based on the day of sacrifice. The experimental group was exposed to ethanol vapours produced in a specially-designed vapour chamber and then compared with age-matched controls. There were 90 eggs in each of the two groups. The development of inter-ventricular septum completed at day 7 of development in chick embryo. Ethanol vapour exposure produced a small discontinuity at day 10 of development in a chick embryo which may be labelled as ventricular septal defect since ventricular development is completed by day7. Interatrial septum formed till day 7 with small perforations which persisted till hatching. Ethanol vapour exposure may lead to ventricular septal defect

developmental gross morphology of pancreas in chick embryo after prenatal administration of valproic acid

To determine the effects of prenatal administration of valproic acid on the developmental gross morphology of pancreas in chick embryo. Experimental study. Anatomy Department, Regional Centre, College of Physicians and Surgeons, Islamabad, from February 2010 to February 2011. An experimental group-A and control group-B, comprised of 30 eggs each. Freshly laid fertilized chicken eggs of experimental group were injected with valproic acid, incubated and hatched. Eggs of control group underwent sham treatment using normal saline. The chicks were sacrificed on hatching or day 22 of incubation, whichever was earlier. The pancreata of only alive chicks of both groups were dissected out, and evaluated for gross morphology in terms of length and weight by statistically comparing with control ones. Then pancreata were stained with aldehyde fuchsin and orange-G stain to study other obvious histological effects, if any. Chicken embryos exposed to valproic acid in ovo, showed significant decrease in length and weight of pancreata. The mean of length [cm] of pancreata in group-A was 2.208 +/- 0.166, and group-B was 2.300 +/- 0.102 [p=0.008]. The mean of weight [g] of pancreata in group-A was 0.032 +/- 0.009, and group-B was 0.048 +/- 0.005 [p=0.001]. Valproic acid exposure showed retarding effect on the gross development of pancreas as depicted by decrease in the length and weight of pancreata

Histomorphometric study of effects of bicalutamide on spermatogenesis in male rats

To study the effects of Bicalutamide on spermatogenesis in male rats. Laboratory based randomized controlled trial. Anatomy Department, Armed Forces Postgraduate Medical Institute [AFPGMI], Rawalpindi in collaboration with National Veterinary Laboratories [NVL], Islamabad from April 2008 to May 2008. Forty adult male Sprague Dawley rats weighing 200-300 grams were randomly divided into two groups, Group A and Group B, each consisting of 20 animals each. Group A was taken as control group and was administered 5 cc of distilled water orally daily for 24 days while group B [Experimental group] was given 5 cc of distilled water daily containing bicalutamide 10 mg/ kg/ day for 24 days. All the animals were sacrificed on the next day after the last dose. The testes were removed and fixed in 10% formalin and then processed for paraffin embedding. Five micron thick sections were made. Haematoxylin, Eosin and PAS stains were used. Histomorphometric analysis was done and parameters, including the tubular diameter, height of seminiferous epithelium and germ cell count were noted. Statistically significant differences were found in tubular diameter, height of seminiferous epithelium and germ cell count in testes of experimental group when compared with the control group. The results showed that the mean tubular diameter, the height of the germinal epithelium of the seminiferous tubules and the number of germ cells were significantly reduced in by the experimental group showing that bicalutamide suppresses spermatogenesis in the Sprague - Dawley rats

Histomorphology of umbilical vessels at regions of low and high altitudes in Pakistan

Objective: To compare the kistomorphological characteristics of human umbilical cord ar high altitde [HA] area i.e. Skardu with those at low altillrde [LA] i.e. Rawalpindi in Pakistan. Materials and methods: This cross sectional cornparalive study was conducted in Anatomy department at Army Medical Gollcgt Rawalpindi, from October 2002 to April 2003. Eighty umbilical cords, forty each from normal cases a: LA and HA! were Included. The full depth samples of umbilcal cord were taken and stained wtth H and E. Dclailed hisromarphology of umbilical cord was studied and thickness of vessel walls via5 measured. Results: Mean thickness ol umbilical artcrics in two gmups was 630.73 5 10.27 Llrn and 534.58 c 8.17 prr., while r-rlcan ihickness of urnbilical vein in same groups was 365.55 + 10.49 urn arid 318.3+ 8.64 Jim respotlively. Tne qlianlitative difference belwccn mean lhickness of unibilical artcries and veins, in LA and HA grnups was statistically significant. Conclusion: Tht thickness d umbilical cord as well as umbilical vessels, was signi1ican:ly less in HA group as compared to LA group.

Effects of free and restricted remobilization on the bodyweight and mid thigh circumference of rabbit

To find out the effects of free and restricted remobilization on the generalized body weight and mid thigh circumference of rabbit. For this purpose, knee joints of rabbits were immobilized first. Randomized Controlled Trials [RCT] Place and Duration of the study: Study was carried out in Army Medical College, Rawalpindi and National Institute of Health Sciences, Islamabad from April 2006 to April 2007. Materials and Twenty four adult New Zealand White rabbits were randomly divided into three groups using random numbers table. Group I served as a control group whereas the left knee joint along with ankle joint of the experimental groups II and III were immobilized in extension in a plaster of Paris cast for 4 weeks. At the end of 4 weeks plaster cast was removed and group II i.e. free mobility group and group III i.e. restricted mobility group [caged animals] were remobilized for another 4 weeks. The generalized body weight and mid thigh circumference of all the animals were taken at the start of experiment, at the end of 4 weeks of immobilization and at the end of 4 weeks of remobilization in all groups. Immobilization caused significant reduction in the body weight and mid thigh circumference of both the experimental groups. On remobilization the body weight increased but significant difference remained between the experimental and control groups. Although weight gain is more in the caged animals during remobilization phase but statistically insignificant difference was found when free and restricted mobility groups were compared. The difference in the mean mid thigh circumference between control and free mobility groups was statistically insignificant whereas the difference between control and restricted mobility groups was significant at the end of 4 weeks of remobilization. Recovery in caged animals was slower as compared to free mobility group in terms of mid thigh circumference but when these two groups were compared statistically insignificant difference existed at the end of 8 weeks. Immobilization induced reduction in body weight and mid thigh circumference which is to a great extent reversible in both free and restricted mobility groups. Both the groups responded almost equally to immobilization stress and confinement did not prevent restoration of bodyweight and mid thigh circumference, however complete recovery of the body weight and mid thigh circumference to initial control values did not occur in 4 weeks

histological study of bowman's glands in human olfactory mucosa

The histology of olfactory mucosa has been previously studied under light and electron microscope. There are marked geographical differences between Pakistan and other countries where most of the research on olfactory epithelium has been conducted. To study morphology and quantitative analysis of Bowman's glands in human olfactory mucosa in Pakistani population. An observational study. This research was done in the Anatomy Department, A M College, Rawalpindi. The duration of study was two years from January 2001 to December 2003. Enbloc specimens were obtained from 20 autopsy cases. After decalcification, they were processed, stained with haematoxylin and eosin [H and E] and seen under light microscope. The olfactory mucosa was observed in the roof, medial and lateral walls of both nasal cavities. The type of glandular tissue and its morphology was observed. The olfactory epithelium was morphologically pseudostraified columnar with a characteristic lamina propria containing numerous olfactory nerve fascicles and Bowman's glands, observed in the roof, medial and lateral walls of both nasal cavities. The secretory acini were almost circular in cross section and measured 20 to 25 micro m in diameter. The secretory cells [7-10 micro m] were pyramidal in shape, with rounded darkly stained nuclei lying in the basal half of the cells. The ducts were seen leading from the glands onto epithelial surface. Mean number of serous acini when compared in the roof, medial and lateral walls of right and left nasal cavities was statistically insignificant. Olfactory mucosa was lined with pseudostratified columnar epithelium. The secretory acini of Bowman's glands were almost circular in cross section and cells were pyramidal in shape. Mean number of serous acini when compared in the roof, medial and lateral walls of right and left nasal cavities was statistically insignificant

comparative histological study of the surface epithelium and high endothelial venules in the subepithelial compartments of human nasopharyngeal and palatine tonsils

To compare the thickness and organization of surface epithelium, and high endothelial venules in subepithelial compartments of human nasopharyngeal and palatine tonsils, with reference to functional differences. Comparative cross-sectional. The Anatomy Department of CPSP Regional Centre, Islamabad, from January to December 2005. Thirty samples each of human nasopharyngeal, right palatine and left palatine tonsils were collected by convenience sampling technique. Haematoxylin and eosin stained paraffin sections were examined for surface epithelium. Thickness of stratified squamous and pseudostratified ciliated columnar epithelium was measured, while organization was observed in case of stratified squamous. The high endothelial venules in subepithelial lymphoid compartments were counted. The surface epithelium of nasopharyngeal tonsils [stratified squamous and pseudostratified columnar inclusive] was 63.21 +/- 1.93 micro m, and that of palatine [stratified squamous] was 143.99 +/- 5.94 micro m thick [p<0.001]. The mean count of high endothelial venules in subepithelial compartments of nasopharyngeal was 1.15 +/- 0.06 and that of palatine tonsil was 0.93 +/- 0.08 [p=0.042]. Organization of stratified squamous epithelium was poor in 26 out of 30 nasopharyngeal, and well in all palatine tonsils [p <0.001]. The surface epithelium of nasopharyngeal tonsil being thinner and poorly organized than that of palatine tonsil might act as a less effective barrier between the antigenic stimulus and subepithelial lymphoid compartments. This may contribute towards higher level of immune response by these compartments of the former, which is endorsed by higher number of high endothelial venules as compared to the latter

Endocrinological response of rabbit after exposure of ovaries to diagnostic ultrasound

To study the serum progesterone and estradiol level in rabbit after exposed of their ovaries to ultrasound waves. Experimental study. NIH, Islamabad, Army Medical College Rawalpindi from Mar 2006 to Sep 2006. Forty, adult, non pregnant female rabbits [Oryctolagus cuniculus strain] weighing 900 gm to 1500 gm were used. They were divided in four groups, each containing ten rabbits, Group I was taken as control. Ovaries of other three groups [group II, III and IV] were exposed to ultrasound [frequency of 3.5 MHz and intensity of 96 dB for two minutes] for 3, 6 and 12 days respectively. Two milliliter of blood samples was withdrawn after 24 hours of last exposure. Progesterone and Estrogen levels in serum were analyzed by Chemiluminescence's method. Initial serum Progesterone and Estrogen levels in rabbits not exposed to ultrasound were compared with ultrasound exposed serum levels. The estradiol level decreased significantly in group IV [P= 0.014] and progesterone level increased significantly in group III [P=0.011]. The current study has revealed that exposure to ultrasound produces changes in hormonal level, if used for longer duration

Histological comparison of human thymus glands between groups of different ages

To compare the thickness of the capsule and interlobular connective tissue, and number and diameter of Hassal's corpuscles in human thymus between two groups of young and old patients. Comparative study. The study was carried out in the Anatomy Department at Army Medical College Rawalpindi from Sep 2007 to Oct 2007. Forty specimens of human thymus were separated into two groups. Group A included 20 specimens from patients between 1-25 years while Group B had same number of specimens of more than 40 years of age. These specimens were fixed in 10% formalin solution and then processed for paraffin embedding. Five micron thick sections were made. Haematoxylin and eosin stain, and PAS stain were used. The thickness of thymic capsule and interlobular connective tissue, number and diameter of Hassal's corpuscles in both groups were noted. Statistically significant differences were found in the thickness of capsule and interlobular connective tissue, number and size of Hassal's corpuscles in specimens of different age groups. In old age, there is definite increase in the thicknesses of capsule and interlobular connective tissue with Hassal's corpuscles decreasing in number but increasing in diameter

Effect of ethanol vapour exposure on survival of chick embryos

To determine the survival of chick embryos after ethanol vapour exposure by noting the number of dead and alive embryos and comparing with age-matched controls. Study Design: Experimental study. Place and Duration of Study: Department of Anatomy at the regional centre of College of Physicians and Surgeons, Islamabad, from February 2006 to February 2007. Chicken eggs, placed in an incubator, were exposed to ethanol vapours produced through a specially designed vapour chamber. The experimental group was dissected on day 7, day 10 and day 22 or hatching whichever was earlier and compared with age-matched controls. The proportion of ethanol vapours in the incubator was monitored with a breathalyzer. No statistical difference was seen in the survival of day 7 alcohol-exposed embryos and their age-matched controls. The survival of day 10-control embryos was significantly higher than alcohol exposed group of same age. The embryos exposed to ethanol vapours from day 1 to day 10 and then followed till hatching or day 22, whichever was earlier, had significantly lower survival than age-matched controls. In this study, ethanol vapour exposure decreased embryo survival with increasing embryonic age and increased duration of exposure

Histological study of human placenta in ultrasonically determined cases of intrauterine growth retardation

To study the light microscopic structure of human placenta in ultrasonically determined cases of intrauterine growth retardation [IUGR]. A cross sectional comparative study. This study was carried out at military hospital Rawalpindi from Jan 2002 to June 2002. Ten placenta of normal and 30 placentae of known intrauterine growth retardation cases were used in this cross sectional comparative study. Placentae were weighed and cut along their maximum diameter into two halves after trimming the membranes. Three specimens were taken: one from the center [A], one from the peripheral margin [C] and one from midway between the two [B]. Specimens were further processed for paraffin sections, 5micro m thick sections were made on rotary microtome. Haematoxylin and eosin [H and E], periodic acid schiff [PAS] and Masson's trichrome stains were used. The morphology of villi was observed and syncytial knots and capillaries were counted. In comprehensive study of the gross observations of the 30 placentae of IUGR cases, it was noted that all [100%] had meconium staining with presence of marginal or retroplacental hemorrhages. Calcification was noted in 24 cases. In the control group mean number of capillaries in A, B and C regions were 114 +/- 14.56, 89 +/- 8.61 and 92 +/- 11.63 respectively. In the IUGR group mean number of capillaries in A, B and C regions were 127 +/- 6.12, 125 +/- 5.53, 122 +/- 7.16 respectively. The difference between mean number of capillaries per field in A, B and C region of control and IUGR group was significant [P<0.05]. Mean birth weight, placental weight, placental diameter and placental thickness in IUGR group was 2.7 +/- 0.200, 163 +/- 18.26, 12.8 +/- 1.18 and 1.46 +/- 0.104 respectively. Difference between placental weight diameter and thickness of normal and IUGR group was statistically significant [p<0.05]. In control group mean number of syncytial knots in A, B and C regions was 114 +/- 7.13, 93 +/- 8.44 and 93 +/- 6.80 respectively. IN IUGR group, the mean number of syncytial knots in A, B and C regions was 169 +/- 7.09, 169 +/- 8.93 and 165 +/- 44.36 respectively. These differences were statistically significant [P<0.05] In terminal villi, syncytial knots and capillaries of the IUGR cases were more in the central region as compared to in the peripheral region. The quantitative difference between syncytial knots and capillaries in IUGR and control group were statistically significant [p<0.05]

Histomorphology of terminal villi of human placenta at low and high altitude with emphasis on blood capillaries

This cross sectional study is designed to compare the histomorphological characteristics of human placenta at high altitude [HA] i.e. Skardu [8500 ft] with those at low altitude [LA] i.e. Rawalpindi [1800 ft]. Cross sectional comparative study. Place and duration of study: The study was conducted in the anatomy department of Army Medical College Rawalpindi, from October 2002 to April 2003. Eighty placentae, 40 each from normal cases at LA and HA, were included. Three full depth samples of placental tissue were taken. Histological study was done in Haematoxylin and eosin [H and E] and Periodic acid Schiff [PAS] stained sections. Detailed morphology of terminal villi was studied and number of capillaries was counted. The core of terminal villus in LA group contained few capillaries with considerable amount of stroma while in HA group it contained abundant dilated capillaries with very little stroma. Mean number of capillaries in LA and HA groups were 201.45 +/- 2.692 and 216.97 +/- 2.828 respectively. The capillaries in terminal villi were dilated and there number was significantly more in high altitude group probably showing adaptive changes in response to hypoxia

Effect of passive tobacco smoking on fertility of female mice

To determine the adverse effects of passive tobacco smoking on fertility of female mice and the preventive role of antioxidants, if any Randomized controlled trial. Anatomy Department, CPSP Regional Centre, Islamabad, from February to July 2005. One hundred and seventeen female mice [Balb C] were selected by random sampling. They were mated and grouped as C [n=30] control, S [n=40] exposed to passive smoke in a whole body exposure chamber and SV [n=37] exposed to smoke and given antioxidants [vitamin C, E]. At 19 days postcoital they were sacrificed and the number of pregnant animals, fetuses and resorption cases were counted. Histological study of uteri without fetuses was done in H and E stained sections for confirmation of pregnancy. Percentages were calculated and Chi-square test was used to calculate statistical significance. The percentage of pregnancies was 55.00% in S and 80.00% in C [p=0.029]. The percentage of animals with more than 11 fetuses was 5.0% in S and 33.30% in C [p=0.001]. The percentage of animals with resorption was 31.80% in S and 0.00% in C [p=0.005]. These values in SV were 64.86%, 18.90% and 20.80%, which were not significantly different from S [p=0.378, 0.216, 0.390 respectively]. Histological study of resorption sites revealed decidual reaction/ remnants of the placenta. Passive tobacco smoke has adverse effects on fertility of female mice, which were not prevented by antioxidants. Either those were due to other chemicals present in smoke, or the antioxidants were inadequate to neutralize the free radicals

Effect of propranolol on prolactin in adult male rats

The study was aimed to observe the effect on blood serum level of prolactin in propranolol treated adult male rats. A experimental study. The study was done at Army Medical College Rawalpindi, National Institute of Health Sciences and NORI, Islamabad from Jan to Aug 2005. Adult male rats were used in the study. Rats were divided into four groups A [Control] and B, C and D [Treatment] each containing fourteen rats. Propranolol 1 mg, 2 mg and 4 mg/kg body weight intraperitoneally daily for sixty days was given to treated groups respectively and control group with 0.5 ml of distilled water. The blood serum level of prolactin was measured in half of the rats of all groups by Enzyme Immunoassay Method. Half of the rats were left for recovery to take place for another sixty days. The above procedure was adopted for the measurement of the prolactin level in the recovered rats. One way ANOVA was used to analyze the data. The prolactin level in control and treated groups rat were 4.75 +/- 1.4 ng/ml serum, 4.70 +/- 1.1 ng/ml serum, 4.51 +/- 1.7 ng/ml serum and 3.1 +/- 0.8 ng/ml serum respectively. Prolactin level was significantly low [p < 0.05] in 4 mg treated groups when compared with control, treated 1 mg and 2 mg groups. After recovery period of sixty days insignificant difference in prolactin level between control and recovered groups was observed. Pattern of low prolactin level was observed following propranolol treatment in male rats

Effects of free mobility verses restricted mobility on the degenerative changes induced by immobilization on the femoral articular cartilage of Rabbit knee

To produce degenerative changes by immobilization of rabbit Stifle [knee] joint in extension and then to see the subsequent effects of either free or restricted mobility on the femoral articular cartilage. An experimental study. This study was conducted from April 2006 to April 2007 at Army Medical College, Rawalpindi and National Institute of Health Sciences, Islamabad. In this study forty adult New Zealand white rabbits were divided into five groups. Groups I and II served as control and were euthanized at 4 and 8 weeks respectively. The left knee joints of the three study groups [III, IV and V] were immobilized in extension by plaster of Paris cast for 4 weeks. After the removal of cast free [IV] and restricted mobility [V] groups were remobilized for another 4 weeks. Following euthanasia, femoral articular cartilages were dissected and examined both macroscopically and microscopically. To evaluate the degenerative changes thickness of articular cartilage, chondrocytes number and semi quantitative Mankin's scoring was done. Immobilization in extension for four weeks produced significant degenerative changes that were evident on both gross and microscopic analysis. Animals in free mobility group showed surface irregularities, superficial ulceration, fibrillation and deep clefts reaching radial zone along with reduction in the staining of matrix. Restricted mobility group when compared with the free mobility group showed significant decrease in Mankin's score and also significant increase in cellularity. The thickness of the articular cartilage in restricted mobility group was although greater than free mobility group but statistically significant difference was not found [p value=.055]. Most peculiar feature of restricted mobility group was proliferation of chondrocytes with formation of clones and intense metachromasia around the clones, a tendency towards intrinsic repair. Early motion and free mobility exposed the injured articular cartilage to increased loads when normal activity was allowed. Moreover rest and restricted mobility for 4 weeks following immobilization allowed some restoration of structure of articular cartilage

Effects of ultrasound on liver

The unicellular organisms and small fish were killed when Ultrasound was first used. Exposure of Rat's liver to varying ultrasound frequencies and time intervals had produced damage ranging from perilobular necrosis to centrilobular damage with lymphocytic infiltration. Keeping in view the growing application trend of diagnostic ultrasound in medicine, its effect's on rabbit's liver were studied. Fifty rabbits with 6501350 gm gross body weight, were studied in five groups. Group I was control and Groups II-V were exposed to an ultrasound frequency of 5 MHz, intensity 100db for 5 minutes daily for 5,10,15 and 30 days. On completion of each exposure, livers were dissected out and examined macroscopically. Macroscopically liver revealed subcapsular hemorrhages and 1-3 mm size connective tissue nodules with solid walls and soft necrotic center. The tissues were processed and stained by heamotoxyllin, eosin and silver impregnation method. The microscopic examination revealed cavitational spaces [dilated sinuses], swollen hepatocytes with large clear spaces, lymphocytic infiltration, perilobular [triad], centrilobular distortion progressing in intensity in groups II to IV. Maximum distortion with necrosis and fibrosis was noted in group V. The findings of the current study reveal that ultrasound may have side effects if used repeatedly especially when for longer duration

Histological study of extrahepatic part of human biliary tree with emphasis on peribiliary glands

Extrahepatic part of human biliary tree was lined with simple tall columnar epithelium. Supranuclear cytoplasm of the cell was darkly stained. Histochemistry of the epithelium revealed that it was mucus secreting. Below the epithelium there was a thick compact layer of connective tissue. Outside the compact layer there was a loose layer of areolar tissue in which smooth muscle, blood vessels and nerves were found. Glandular acini were present within and outside the compact layer of intrapancreatic bile duct [IPBD] and common bile duct [CBD] whereas these were present only within the compact layer of bile duct of ampulla of Vater [BDAV]. Statistical significance of quantitative difference between glandular acini of BDAV with other regions revealed that the difference was significant [P < 0.01] with IPBD and non significant [P > 0.05] with CBD. Statistical significance of quantitative difference between glandular acini of IPBD with other regions revealed that the difference was significant [P < 0.01 and P < 0.05] with BDAV and CBD respectively. Statistical significance of quantitative difference between glandular acini of CBD with other regions also revealed that difference was significant [P<0.05] with IPBD whereas it was not significant with BDAV [P>0.05]. Intramural and extramural glandular acini were present in IPBD and CBD whereas BDAV contained only intramural glands. The difference between intramural and extramural glandular acini was statistically significant [P < 0.02] in IPBD. The difference between intramural and extramural glandular acini of CBD was also statistically highly significant [P<0.001]. Glandular acini as demonstrated by different stains were 100% mucous type in BDAV and IPBD whereas it was mixed type in CBD. The acini of CBD were serous, mucous and mixed type. The difference between serous and mucous, mucous and mixed glandular acini was statistically highly significant [P<0.001]. The difference between serous and mixed glandular acini was statistically not significant [P>0.05]. Ratio of serous, mucous and mixed glandular acini was 1:22:1 respectively